Objective The experiment aims at probing the best condition of the isolation and purification of rat islets.Methods The islets were isolated from rat pancreas by hepatopancreatic duct perfusing with collagenase and purified with Ficoll 400 discontinuous density gradient centrifugation.Then the purified islets were subjected to histological staining,electron microscopy and radioimmunoassay for identification of specificity and viability.Results The histological staining revealed that the viability and the purity of the purified islets were above 95％and 85％respectively.Electron microscopy showed that the purified islets were morphologically intact with clear membrane and plenty of secreting granules.Radioimmunoassay demonstrated the secreted insulin concerntration between low-glucose groups and high-glucose groups varied significantly,which verified the good function of the islets.Conclusion Hepatopancreatic duct perfusing with collagenase is a good method for digestion.There aye many factors that influence the quantity and quality of the acquired islets,such as the completed expansion of pancreas,the concentration and viability of collagenase and the digested time,etc.

Objective The experiment aims at probing the best condition of the isolation and purification of rat islets. Methods The islets were isolated from rat pancreas by hepatopancreatic duct perfusing with collagenase and purified with Ficoll 400 discontinuous density gradient centrifugation.Then the purified islets were subjected to histological staining,electron microscopy and radioimmunoassay for identification of specificity and viability. Results The histological staining revealed that the viability and the purity of the purified islets were above 95% and 85% respectively.Electron microscopy showed that the purified islets were morphologically intact with clear membrane and plenty of secreting granules.Radioimmunoassay demonstrated the secreted insulin concerntration between low-glucose groups and high-glucose groups varied significantly,which verified the good function of the islets.Conclusion Hepatopancreatic duct perfusing with collagenase is a good method for digestion.There are many factors that influence the quantity and quality of the acquired islets,such as the completed expansion of pancreas,the concentration and viability of collagenase and the digested time,etc.

Objective To understand the mechanism of how cardiomyocytes exit from the cell cycle,we examined the expression of polo-like kinase 1(plk1) in the postnatal developmental process of cardiac myocytes. Methods Mitotic Index(MI) of cardiomyocytes was examined in the neonatal,2-week-old,4-week-old,and adult rat hearts(five cases per groups) by double immunofluorescence stained with H3P and ?-sarcomeric actin antibodies.plk1 mRNA and protein expression during the postnatal developmental process of cardiac myocytes were detected by RT-PCR and Western blot analysis in rat hearts. Results The MI of cardiomyocytes in 0-day-old hearts(0.905?0.087%) was approximately 2.4 times over that in 2-week-old hearts(0.372?0.094%)(P

Objective To investigate the activity changes of gelatinase in the formation of ascending aortic aneurysm.Methods Thirty five young Wistar rats were divided into two groups:the control group and the experiment group.The rat models induced by ascending aorta banding were made.The ascending aortas were taken after 3-5 months operation,changes of gelatinase activity was observed by gelatin zymography and film in situ zymography.Results Gelatinase activity of ascending aortic aneurysm was significantly increased compared with that of normal ascending aortic aorta.Conclusion Elevation of gelatinase activity may play a significant role in the formation of ascending aortic aneurysm.

Objective To investigate the expressions and significance of transforming growth factor-beta 1(TGF?1) and its typeⅡ receptor(TGF?RⅡ) in experimental rat ascending aortic aneurysm of rat model.Methods The rat ascending aortic aneurysm models were made by banding ascending aorta of young Wistar rats.The ascending aortas were taken 4 months after operation.Immunohistochemistry staining and Western blotting were used to investigate the expressions of TGF?1 and TGF?RⅡ.Result Immunohistochemistry staining results showed that TGF?1 expressed in all layers of the aortic aneurysm and the control.TGF?RⅡ was extensively located in the hyperplastic intima and tunica media smooth muscle cells in the aortic aneurysm,while there was only a little positive staining in the control group.Western blotting results indicated that the expression levels of TGF?1 and TGF?RⅡ in the aortic aneurysm were stronger than the control,P

Adolescent ; Child ; Child, Preschool ; Ear, Middle ; physiopathology ; Female ; Humans ; Male ; Sleep Apnea, Obstructive ; physiopathology

Objective To evaluate the growing condition of human umbilical vein endothelial cells (HUVECs), which were cultured on the membrane of different component, such as Chinese medicine, before or after the alkali treatment of 3-hydroxybutyrate-co-3-hydroxyhexanoate copolyesters (PHBHHx) and the biocompatibility between PHBHHx flim and endothelial cell. Methods The HUVECs were harvested by Baudine method,and identified by immunohistochemical method.Then the HUVECs of third passage were inoculated on the material surface and cultured for 8 hours,12 hours and 24 hours. After that, the morphology of HUVECs on different surfaces were observed by scanning electron microscopy, the distribution condition on different membranes were compared by cell-labeling immunofluorescence, and the cell viabilities of all groups were detected by MTT method. Results The HUVECs were successfully separated,and immunohistochemistry staining of FLK-1 and factor Ⅷ was positive.The result of HUVECs culture showed that cells on the material surface growed well, and proliferated significantly. The MTT analysis showed that the PHBHHx film of surface modification and adding some certain proportion of Chinese medicine could promote the growth and proliferation of HUVECs in vitro,and the cells were thriving, full shape, distribution on the surface by scanning electron microscopy and fluorescence microscopy.Conclusion The PHBHHx film of surface modification and containing certain proportion of Chinese medicine coating had good compatibility of HUVECs, which was favourable to cell growth, adherence and proliferation in vitro.

Infant hair is a good medium to assess the cumulative exposure level of infant in the mother ' s body. Chlorinated persistent organic pollutants were ubiquitous and with the highest volume concentration in the human body. In this experiment, 20 infant hairs were collected from Beijing. The sample was overnight incubated in HCl, extracted with the mixture of hexane and dichloromethane, and cleaned up by a cartridge filled with alumina and acidified silica. The final eluate was concentrated, and 16 kinds of typical chlorinated persistent organic pollutants were detected with gas chromatography_high resolution mass spectrometry ( HRGC_MS). The results showed that the detection limit of the target compounds in sample was 1. 00-2. 50 μg/kg, the recovery of surrogate in all samples was more than 67 . 6%, and the range of recoveries for target compounds in spiked sample was 62. 5%-92. 3%. The positive rate is 100% for hexachlorobenzene,β_HCH and p, pˊ_DDE, 85% for γ_HCH, 50% for PCB28, and 40% for PCB52. The concentrations of Hexachlorobenzene, HCHs, DDTs and PCBs were 5. 48-8. 40 μg/kg, 3. 86-27. 1 μg/kg, 1. 16-18. 3 μg/kg and 2. 20-22. 1 μg/kg, respectively. The average concentrations were 7. 84 μg/kg for hexachlorobenzene, 6. 93 μg/kg for HCHs, 5. 53 μg/kg for DDTs, and 3. 44 μg/kg for PCBs. The method and the analysis results can be used to evaluate the accumulation level and cumulative exposure level of 16 target compounds for fetal in motherˊs body.

An enantioselective method was developed for the separation and determination of three chiral hexabromocyclododecanes ( HBCDs ) including α-HBCD, β-HBCD, γ-HBCD in soil and earthworm by HPLC-ID-MS/MS. d18-HBCDs used as internal standards were added to the samples before extraction. HBCDs enantiomers were extracted from soil by accelerated solvent extraction ( ASE ) with n-hexane/DCM (1:1,V/V) at 100℃ and 10 MPa for 5 min, and further cleaned up using silica column. HBCDs enantiomers were extracted from earthworm by vortex turbulence with ethyl acetate. The extracts were orderly sulphonated by sulfuric acid, and purified by silica column. For all HBCDs enantiomers, good linearities were obtained in the concentration range of 0. 25-50 ng/mL. Limits of detection ( LOD) and limits of quantification ( LOQ) were 0. 00544-0. 00766 ng/g and 0. 0173-0. 0244 ng/g, respectively in soil. The recoveries of spiked samples at 0. 05 and 2. 5 ng/g levels were 80. 0%-95. 9% with relative standard deviations ( RSD, %) of 5. 7%-11. 9% in soil. Limits of detection (LOD) and limits of quantification (LOQ) were 0. 0103-0. 0148 ng/g and 0. 0328-0. 0471 ng/g, respectively in earthworm. The recoveries of spiked samples at 0. 1 and 5 ng/g levels were 78. 0% -94. 4% with relative standard deviations ( RSD, %) of 6. 1% -12. 2% in earthworm. This method can meet the requirements of determination of trace HBCDs in soil and earthworm.

Objective To assess the consistency of four standardized cystatin C particle-enhanced turbidimetric assay (PETIA) and one particle-enhanced nephelometric immunoassay (PENIA) measurement systems Methods Performance verification test was conducted according to CLSI EP 15-A2 and EP9-A2. Fourty serum samples in comparative test were obtained from the remaining serum samples of outpatients in Peking Union Medical College Hospital in March 2013.Fourty serum samples were tested on Olympus AU5400 automatic biochemical analyzer ( four PETIA Cys C reagents:Shanghai Jingyuan Co ., Ltd, Beijing Leadman Biochemistry Co ., Ltd, Beijing Strong Biotechnologies , Maker Biotechnology in Sichuan , and labelled as A, B, C, D respectively) and PENIA N Latex Cys C measurement system on Siemens BNⅡ(labelled as E).Correlation analysis were performed among four PETIA methods one PENIA method Differences of each detection system were compared in the medical decision level 1,2,3,4 mg/L.The reference material ERM-DA471/IFCC was measured by five systems and bias ( percentage bias ) was calculate for each system.Results Results of systems A, B, C, D, E were 1.29(0.89-2.43), 1.30 (0.96-2.59), 1.22(0.90-2.44), 1.27(0.96-2.47), 1.14(0.82-2.05)mg/L.Chart shows bias among these five systems was small when Cys C concentration was less than 4mg/L.PETIA method A, B, C, D correlated with their mean value well , with the average deviation from their mean value ( percent deviation) at -0.017 -0.031 mg/L ( -3.1%-2.1%), and all were less than allowed bias from the biological variation (3.4%).The deviation of PETIA method A, B, C, D with their mean value in medical decision level at -0.176 -0.178 mg/L.Systems A, B, C, D correlated well with the result of PENIA method system E , and the mean deviation ( percent deviation ) was at 0.278 -0.326 mg/L ( 12.6%-18.5%) , and the deviation ( percent deviation ) in the medical decision level 0.055 -1.079 mg/L (5.51%-26.98%).Bias of PETIA method A, B, C, D Cys C system measuringERM -DA471/IFCC ranged from 0.22 to 0.39 mg/L ( 3.9%-7.0%) , which exceeded the allowable range of the reference material target value, and were larger than the allowable bias from biological variation (3.4%).Bias ( percent ) of PENIA method system E was -0.1 mg/L ( -1.7%) , within the allowable range of ERM-DA471/IFCC target value .Conclusions The consistency of four assesed PETIA Cys C reagents was relatively ideal, and improved markably after being traced to ERM-DA471/IFCC.Besides, the results of PETIA were higher than those of PENIA .Bias among these five systems was small when Cys C concentration was less than 4 mg/L, and the bias became larger in higher Cys C concentration.